LOCUS       sequence-295386-        4962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF8 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4962)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4962)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4134215
FEATURES             Location/Qualifiers
     source          1..4962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             368..388
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             737..754
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    831..952
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(842..861)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     896..915
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1067..1113
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1143..1279
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1324..1395
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1621..2209)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1701..1720)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2955..3023,3024..3815)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3173..3192)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3828..3964
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4116..4187
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4192..4238)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4256..4304
                     /note="synthetic polyadenylation signal"
     enhancer        4385..4688
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4690..4892
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4843..4863
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaaa tttcttgttt tcttaggaat catcacaact gtagctgcat ttcaccaaga
       61 atgtagttta cagtcatgta ctcaacatca accatatgta gttgatgacc cgtgtcctat
      121 tcacttctat tctaaatggt atattagagt aggagctaga aaatcagcac ctttaattga
      181 attgtgcgtg gatgaggctg gttctaaatc acccattcag tacatcgata tcggtaatta
      241 tacagtttcc tgtttacctt ttacaattaa ttgccaggaa cctaaattgg gtagtcttgt
      301 agtgcgttgt tcgttctatg aagacttttt agagtatcat gacgttcgtg ttgttttaga
      361 tttcatcgaa aacctttact ttcagtcgat gagtgacaaa atcattcatc taaccgatga
      421 ctcctttgat acagacgtgt tgaaggccga tggtgcgata ctggtagatt tctgggctga
      481 atggtgtggc ccgtgcaaga tgatcgcacc cattctggac gagatcgccg acgagtatca
      541 aggaaaatta actgtcgcta agcttaatat agatcagaac ccagggacgg cacctaaata
      601 cggcatcagg ggaataccta ccctcctgct ctttaagaat ggggaagttg ccgcgacaaa
      661 agtgggagcc ctgtctaagg gccagctaaa agagttcttg gacgctaacc tggcaggttc
      721 aggctcgggg cacatgcacc atcaccatca ccatagcagc gggctcgtgc cacgcggcta
      781 agctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
      841 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
      901 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
      961 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     1021 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
     1081 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
     1141 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     1201 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     1261 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
     1321 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
     1381 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
     1441 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
     1501 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
     1561 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1621 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1681 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1741 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1801 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1861 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1921 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1981 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2041 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2101 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2161 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2221 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2281 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2341 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2401 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     2461 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     2521 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     2581 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     2641 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     2701 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     2761 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     2821 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     2881 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     2941 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3001 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3061 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3121 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3181 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3241 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     3301 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3361 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3421 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3481 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     3541 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     3601 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3661 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3721 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3781 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     3841 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     3901 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     3961 ttaagatccc gaatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc
     4021 ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt
     4081 cagcttacct ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg
     4141 tatcttcttc cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc
     4201 cctcaagacc cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac
     4261 aaaaaaccaa cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg
     4321 agtgtagcca gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     4381 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     4441 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     4501 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     4561 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     4621 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     4681 ttaccatgct gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     4741 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     4801 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     4861 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt
     4921 tgtcgatcct accatccact cgacacaccc gccagcggcc gc
//
